Leukemias induced by ethylnitrosourea in Wistar rats: incidence and chemotherapy.

After application of 15 mg/kg ethylnitrosourea (ENU) weekly for 15 weeks to 200 male Wistar rats, 76 developed acute leukemias, 15 developed chronic myeloid leukemias and 39 had malignomas in other organs. The weekly application of 75 mg/kg ENU to another group of 200 male Wistar rats for 3 weeks caused 57 acute leukemias, 6 chronic myeloid leukemias and 47 malignomas in other organs. From 112 “autochthonous” acute leukemias which were treated with a combination of vincristine, adriamycin and cytosine arabinoside (ADOA), 67 (60%) achieved a complete remission with a median survival time of 64 days (range 25–173). The survival time of this group was significantly increased compared to untreated controls (α = 0.01). Forty-five rats (40%) with acute leukemia died during the first week after start of therapy, or therapy had to be discontinued. The failure of treatment in this group was attributed to the poor general condition of the animals whose platelet counts and hemoglobin values were low, due to late diagnosis.The “autochthonous” acute leukemias demonstrated some parallels with human acute leukemia. Their use in experimental chemotherapy is discussed.     

白介素-18结合蛋白对阿霉素肾病大鼠Fractalkine表达的影响

目的探讨不同时期阿霉素肾病大鼠尿液中Fractalkine表达的变化及白介素18结合蛋白(IL-18BP)的治疗作用。方法建立阿霉素肾病大鼠模型,设立肾病组(n=12)、IL-18BP干预组(n=11)和对照组(n=10)。观察42 d,定期检测大鼠尿Fractalkine水平及24 h尿蛋白定量。结果肾病组和IL-18BP干预组大鼠尿Fractalkine水平均进行性提高,差异有统计学意义(F=91.395、47.248,P均<0.01);IL-18BP干预组大鼠各时间点尿Fractalkine水平均低于肾病组(P均<0.05),高于对照组(P均<0.01)。肾病组、IL-18BP干预组大鼠尿Fractalkine水平与24 h尿蛋白定量均呈正相关(r=0.970、0.904,P均<0.01)。结论 Fractalkine与阿霉素肾病大鼠蛋白尿相关,IL-18BP可降低Fractalkine水平,可能有助于微小病变肾病治疗。     

Enhanced desensitization efficacy by liposomal conjugation of a specific antigen

Since liposomes are known as strong adjuvants, we attempted to use liposomes in immunotherapy as adjuvants, and to achieve desensitization in pre-sensitized mice. At first, we sensitized mice with intraperitoneal injection of model antigen, 100 microg ovalbumin (OVA), with Alum and treated them with liposome composed of distearoylphosphatidylcholine (DSPC) and cholesterol (2:1 as a molar ratio), which was coupled with a small amount of OVA (10 microg OVA in 400 nmol DSPC and 200 nmol cholesterol-liposome was injected into 20 g mouse). It is well known that antigen-specific immunotherapy increases IgG blocking antibodies and decreases in IgE antibodies. The treatment with i.v. injection of OVA-liposome at days 8, 10, and 12 after sensitization strongly suppressed OVA-specific IgE production without affecting IgG level after the boost (100 microg OVA with Alum). Moreover, the treatment with high-density OVA-liposome (10 microg OVA in 80 nmol DSPC and 40 nmol cholesterol-liposome/20 g mouse) not only strongly suppressed IgE levels but also reduced IgG production after the boost of OVA-sensitized mice suggesting the importance of liposomal characteristic in desensitization immunotherapy. Next we reduced the dose of OVA-liposome and the desensitization effect was also observed at the dose of as low as 1 microg OVA on OVA-liposome/mouse. On the contrary, free OVA did not affect the production of both IgG and IgE levels. Biodistribution study indicated that OVA-liposome was highly accumulated in spleen of OVA-sensitized mice compared to control liposome at 3 h after i.v. injection. These results suggest that the liposomal OVA effectively interacts with and desensitizes immune cells, therefore, liposomes coupling with a certain antigen may be effective in allergy immunotherapy.     

阿霉素中毒大鼠膈肌骨架蛋白和肌浆网钙泵的表达变化及意义

目的观察不同剂量阿霉素诱导大鼠膈肌急性损伤时膈肌收缩特性的变化与大鼠膈肌细胞骨架蛋白和肌浆网钙泵表达变化的关系。方法一次性腹腔注射阿霉素制备急性膈肌损伤动物模型,♂SD大鼠随机分成:阿霉素低剂量组(10mg·kg-1)、中剂量组(20mg·kg-1)和高剂量组(40mg·kg-1)及对照组,每组10只。对照组用生理盐水腹腔注射。3d后剖胸取膈肌,应用体外灌流大鼠膈肌肌条方法,分别测量其单收缩张力(Pt)、最大强直张力(Po)、峰值收缩时间(CT)、半舒张时间(1/2RT),张力最大上升速度(+dt/dtmax)及张力最大下降速度(-dt/dtmax)。应用逆转录聚合酶链反应(RT-PCR)测定膈肌骨架蛋白肌联蛋白(titin)、伴肌动蛋白(nebulin)和肌浆网钙泵(sarcoplasmic reticulum Ca2+-ATPase,SERCA)mRNA的表达。结果与对照组相比,阿霉素中、高剂量模型组大鼠膈肌Pt,Po,±dt/dtmax下降(P<0.01),CT、1/2RT延长(P<0.01)。阿霉素中毒大鼠膈肌各基因表达较对照组相比分别降低(P<0.01)。结论阿霉素中毒大鼠膈肌细胞膈肌骨架蛋白titin、nebulin和SERCA mRNA表达下调,并可能与膈肌张力改变有关。     

高效液相色谱法测定U87细胞内外液中的阿霉素含量

目的:建立U87肿瘤细胞中阿霉素的高效液相色谱荧光检测方法,为进一步研究化疗药对肿瘤细胞功能和活性的影响及逆转耐药研究提供方法学基础。方法:U87细胞以1×106·mL-1密度与不同浓度阿霉素孵育24h为药物处理条件。将细胞分为空白对照组和加药组,采用HPLC法测定细胞内外阿霉素的含量。色谱柱为Inertsil ODS-3(250mm×4.6mm,5μm);流动相为甲醇-乙腈-0.01mol·L-1磷酸二氢铵(40∶15∶45)(以0.1%冰醋酸调整pH至3.52);柱温35℃,流速1.4mL·min-1,荧光检测器检测波长λex=495nm,λem=560nm,以盐酸柔红霉素为内标。结果:细胞内阿霉素在0.025～7.5mg·L-1范围内线性关系好,r=0.9999;细胞外阿霉素在0.1～10.0mg·L-1范围内线性关系好,r=0.9999。细胞内外液阿霉素的检测限均为0.005mg·L-1,细胞外液的定量限为0.1mg·L-1。日内、日间RSD均小于10%,样品稳定性好。结论:采用HPLC荧光法测定U87细胞内外液中阿霉素的含量,方法简单、准确、线形范围宽、灵敏度高、精密度好,可用于对培养的肿瘤细胞内外...     

阿霉素对端粒序列及其相似物电泳迁移速度的影响

目的 观察阿霉素对端粒序列d(TTAGGG)不同重复次数以及与其具有相似结构的单核苷酸序列二级结构的影响,探讨阿霉素诱导端粒复重序列d(TTAGGG)n形成鸟嘌呤四联体的机制.方法 人工化学合成线性单核苷酸序列d(TTAGGG)、d (TTAGGG)2、d(TTAGGG)3、d(TTAGGG)4、d(TTAGGG)5、d(TTGGAG)4、d(TGTAGG)4、d(TAGGTG)、d(TTGAGG)4;用非变性聚丙烯酰胺凝胶电泳法观察不同浓度阿霉素(0、1.25、2.5、5.0μg/mL)对端粒重复序列d(TTAGGG)4电泳迁移速度的影响;观察5.0μg/mL阿霉素对d(TTAGGG)、d(TTAGGG)2、d(TTAGG)3、d(TTAGGG)4、d(TTAGGG)5电泳迁移速度的影响;观察5.0 μg/mL阿霉素对d(TTAGGG)4、d(TTGGAG)4 、d(TGTAGG)4、d(TAGGTG)4 、d(TTGAGG)4电泳迁移速度的影响.结果 随着阿霉素的浓度的提高,端粒重复序列d(TTAGGG)4电泳迁移速度也随之加快;阿霉素可使端粒重复序列d(TTAGGG)4、d(TTAGGG)5序列的二级结构发生改变,从而加快其电泳迁移的速度,对d(TTAGGG)、d(TTAGGG)2、d(TTAGGG)3的电泳迁移速度没有影响;阿霉素可对含有GGG碱基排列的端粒重复序列发生电泳迁移速度的加快,而对无GGG碱基排列的端粒序列类似物的电泳迁移没有影响.结论 在阿霉素存在条件下,端粒序列d(TTAGGG)n重复4次以上的寡核苷酸序列的电泳迁移速度加快,其二级结构的改变是迁移速度加快的主要原因,这种改变后形成的二级结构可能为G-四联体.     

姜黄素对阿霉素肾病大鼠NADPH氧化酶和nephrin表达的影响

目的通过观察姜黄素对阿霉素肾病大鼠肾组织中尼克酰胺腺嘌呤二核苷酸磷酸（NADPH）．氧化酶与肾小球裂隙膜分子nephrin表达的影响,探讨姜黄素治疗肾病综合征的作用机制。方法将Sprague．Dawley大鼠50只随机分为5组（正常对照组、模型组、姜黄素低剂量干预组、姜黄素高剂量干预组和波尼松干预组）。除正常组大鼠外,各组大鼠尾静脉一次性注射阿霉素（5mg／kg）造模。分别在建模后第28天收集24h尿液,第29天处死所有大鼠,取肾脏组织,应用化学比色法检测肾组织氧化应激指标丙二醛（MDA）和超氧化物歧化酶（SOD）,实时荧光定量聚合酶链式反应法和蛋白质印迹法检测肾组织NADPH氧化酶活性的关键亚基p47phox和neprhinmRNA与蛋白的表达变化。结果与正常对照组相比,模型组大鼠24h尿蛋白量、肾组织MDA含量明显增加,肾组织p47phoxmRNA和蛋白表达明显增加,SOD活性明显下降;肾组织中nephrinmRNA和蛋白表达均明显减少。与模型组比较,姜黄素干预组尿蛋白、肾组织MDA明显减少,SOD活性明显增加。姜黄素干预组可显著下调p47phoxmRNA和蛋白表达;姜黄素干预组nephrinmRNA和蛋白表达均明显增加。结论姜黄素能减轻肾小球滤过屏障损伤,降低尿蛋白,抑制肾组织p47ph“表达和增加nephrin表达可能是其作用机制之一。     

In vitro and in vivo evaluation of targeting efficiency of Adriamycin hydrochloride magnetic microspheres

The objective of this study was to prepare magnetic microspheres as a targeting drug delivery system and to specifically evaluate its targeting efficiency. The magnetic microspheres were prepared by emulsion cross-linking techniques. Targeting efficiency was specifically investigated by experiments of biodistribution on rats and histological study. Adriamycin hydrochloride (ADR)-loaded magnetic microspheres were successfully prepared with the mean diameter of 3.853 μm (± 1.484 μm), and had its speciality of superparamagnetism. The results of the targeting efficiency study showed that application of the external magnetic field significantly increased the ADR concentration from 40.28 μg/ml to 100.70 μg/ml at 10?min, 36.99 μg/ml to 91.16 μg/ml at 60?min, and 13.71 μg/ml to 28.30 μg/ml at 180?min in liver as the targeting tissue. The relative uptake efficiencies in liver by injection treatment of ADR magnetic microspheres with external magnetic field were 3.87, 5.59, and 3.34 at 10?min, 60?min, and 180?min after administration, respectively. In conclusion, distinguished targeting efficiency was displayed, which indicated that the magnetic microspheres could be applied as a novel targeting drug delivery system.     

芪蝼肾炎丸对阿霉素大鼠蛋白尿及肾脏ET-1及TGF-β1的影响

目的：观察芪蝼肾炎丸对阿霉素大鼠尿蛋白、血清生化指标以及肾脏组织内皮素1（ET-1）、转化生长因子-β1（TGF-β1）的影响。方法制备阿霉素肾病大鼠模型,检测大鼠24h尿蛋白定量,血清生化,肾脏ET-1、 TGF-β1的变化。结果芪蝼肾炎丸可明显降低阿霉素大鼠24h尿蛋白定量、血浆胆固醇、三酰甘油,升高血浆白蛋白,并使肾组织的ET-1及TGF-β1水平下降。结论芪蝼肾炎丸可降低阿霉素大鼠的血脂和蛋白尿水平,升高血清白蛋白,其机理与下调肾组织ET-1、 TGF-β1的水平有关。